Compositions and methods for deep dermal drug delivery

ABSTRACT

Pharmaceutical compositions for the topical administration of a drug to the pilosebaceous unit and methods for administering the same. As disclosed herein, the inventors of the present invention have made the surprising discovery that pharmaceutical compositions comprising small particles of an active pharmaceutical ingredient can be administered to the pilosebaceous unit. The pharmaceutical composition can comprise SHR0302 or spironolactone as an active pharmaceutical ingredient.

CROSS REFERENCE TO RELATED APPLICATIONS

The present application claims priority to U.S. Provisional ApplicationNo. 63/114,887 filed on Nov. 17, 2020 and U.S. Provisional ApplicationNo. 63/221,349 filed on Jul. 13, 2021, the disclosures of which areincorporated herein in their entirety by reference.

FIELD OF THE INVENTION

The subject matter disclosed herein generally relates to pharmaceuticalcompositions for the topical administration of a drug to thepilosebaceous unit and methods for administering the same. As disclosedherein, the inventors of the present invention have made the surprisingdiscovery that pharmaceutical compositions comprising small particles ofan active pharmaceutical ingredient can be targeted to the pilosebaceousunit. In preferred embodiments, the pharmaceutical composition comprisesSHR0302 or spironolactone as an active pharmaceutical ingredient.

BACKGROUND OF THE INVENTION

Transdermal and topical delivery of drugs have a variety of advantagescompared with other routes of administration. Transdermal and topicaldelivery can be used to deliver drugs continuously into the systemiccirculation and circumvent first-pass metabolism. In contrast, there isa significant first-pass effect of the liver that can prematurelymetabolize drugs for oral drug delivery. Transdermal and topicaldelivery also have advantages over intravenous administration, whichmust be sterile products and can be painful thereby increasingnoncompliance by patients. Transdermal delivery on the other hand can benon-sterile, non-invasive and self-administered.

Traditional drug delivery systems have focused on administration via thetransepidermal route of delivery. Andrea C. Lauer et al.,Transfollicular Drug Delivery, Pharmaceutical Research 12:2 (1995). Theskin consists primarily of four layers: (a) the stratum corneum(nonviable epidermis), (b) viable epidermis, (c) dermis, and (d)subcutaneous tissues. The skin also contains appendages in the form ofterminal hairs, which may extend more than 3 mm below the skin surfaceinto the subcutaneous fatty tissue and vellus hair, which is the fine,often unnoticed body hair that extends less than 1 mm into the dermis.The hair follicle, hair shaft, and sebaceous gland, which secrets alubricating oil matter into the hair follicles, comprise what is knownas a pilosebaceous unit. While the stratum corneum has traditionallybeen viewed as the primary pathway for the penetration of drugs, it isalso the main barrier to percutaneous absorption. In the past,researchers have questioned the significance of the pilosebaceous unitin drug delivery.

More recently, however, the potential role of the pilosebaceous unit andalternative mechanisms for the transdermal delivery of drugs have beeninvestigated. Amit Verma et al., Transfollicular drug delivery: currentperspectives, Research and Reports in Transdermal Drug Delivery (Apr.20, 2016). The mammalian hair follicle is a complex, dynamic structurein which unique biochemical and immunological reactions occur. While thepilosebaceous unit may be an acceptable target for drug delivery, thereare several challenges to drug delivery to the pilosebaceous unit. Oneof the challenges relating to drug delivery to the pilosebaceous unit isthe need to bypass the stratum corneum, which extends approximately10-20 μm deep and the upper capillary plexus, which extendsapproximately 80 μm deep.

There is currently a need for pharmaceutical compositions capable ofpenetrating deeper into the dermis, approximately 1,000 μm to 2,000 μmto the pilosebaceous unit. There is an unmet need for novelpharmaceutical compositions and methods of administering drugs via thepilosebaceous unit.

SUMMARY OF THE INVENTION

The present invention relates to pharmaceutical compositions for thetopical administration of a drug to the pilosebaceous unit and methodsfor administering the same. The inventors of the present invention havemade the surprising discovery that a pharmaceutical compositioncomprising small particles of an active pharmaceutical ingredient can bedelivered to the pilosebaceous unit resulting in deeper penetration intothe dermis and improved efficacy. In preferred embodiments, thepharmaceutical composition comprises SHR0302 or spironolactone as anactive pharmaceutical ingredient.

In certain embodiments of the present invention, a pharmaceuticalcomposition is provided comprising a therapeutically effective amount ofan active pharmaceutical ingredient and a silicone selected from thegroup consisting of dimethicone, cyclomethicone, and combinationsthereof. The active pharmaceutical ingredient has a primary particlesize distribution that is characterized by a D90 value of less thanabout 20 μm. In certain embodiments, the pharmaceutical ingredient has aprimary particle size distribution that is characterized by a D90 valueof less than about 10 μm or more preferably less than about 5 μm. Incertain embodiments, the composition comprises about 0.10% w/w to about7.5% w/w of the active pharmaceutical ingredient. The pharmaceuticalcompositions of the present invention can be capable of delivering theactive pharmaceutical ingredient to the pilosebaceous unit. In certainembodiments, the active pharmaceutical ingredient is capable ofachieving dermal penetration of at least 1 mm in the subject.

In certain embodiments of the present invention, a pharmaceuticalcomposition is provided comprising a therapeutically effective amount ofSHR0302 or a pharmaceutically acceptable salt thereof. The SHR0302 canhave a primary particle size distribution characterized by a D90 valueof less than about 20 μm, less than about 10 μm, or more preferably lessthan about 5 μm. The SHR0302 can further have a primary particle sizedistribution characterized by a D50 value of less than about 5 μm, lessthan about 1 μm, or more preferably less than about 0.7 μm. The SHR0302can further have a primary particle size distribution characterized by aD10 value of less than about 1 μm, less than about 0.5 μm, or morepreferably less than about 0.25 μm.

The pharmaceutical composition further comprises a silicone selectedfrom the group consisting of dimethicone, cyclomethicone, andcombinations thereof. In certain embodiments of the present invention,the pharmaceutical composition comprises SHR0302 suspended in at leastone of dimethicone and cyclomethicone.

In certain embodiments, the pharmaceutical composition can compriseabout 0.10% w/w to about 5% w/w of SHR0302 or a salt thereof. Inpreferred embodiments, the pharmaceutical composition can comprise fromabout 0.1% w/w to about 3% w/w of SHR0302 or a salt thereof.

In certain embodiments of the present invention, a method of treatingalopecia areata in a subject in need thereof is provided. The methodcomprises topically administering to the subject the pharmaceuticalcompositions of SHR0302 described herein. In the methods of the presentinvention, the SHR0302 or salt thereof can be delivered to thepilosebaceous unit. In the methods disclosed herein, administration ofthe pharmaceutical composition can result in dermal penetration ofSHR0302 of at least about 1 mm in the subject, and preferably to thedepth of the hair bulb for a terminal hair, about 2 to 3 mm in thesubject.

In certain embodiments of the present invention, a pharmaceuticalcomposition is provided comprising a therapeutically effective amount ofspironolactone or a pharmaceutically acceptable salt thereof. Thespironolactone can have a primary particle size distributioncharacterized by a D90 value of less than about 6 μm, less than about 1μm, or more preferably less than about 0.25 μm. The spironolactone canfurther have a primary particle size distribution characterized by a D50value of less than about 2.7 μm, less than about 0.75 μm, or morepreferably less than about 0.15 μm. The spironolactone can further havea primary particle size distribution characterized by a D10 value ofless than about 1.2 μm, less than about 0.50 μm, or more preferably lessthan about 0.10 μm.

The pharmaceutical composition further comprises a silicone selectedfrom the group consisting of dimethicone and cyclomethicone. In certainembodiments of the present invention, the pharmaceutical composition ofspironolactone is an oil-in-water emulsion.

Further, the pharmaceutical composition can comprise about 0.10% w/w toabout 7.5% w/w of spironolactone or a salt thereof. In certainembodiments, the pharmaceutical composition can comprise from about 0.5%w/w to about 5% w/w of spironolactone or a salt thereof.

In certain embodiments of the present invention, a pharmaceuticalcomposition is provided comprising a therapeutically effective amount ofspironolactone or a pharmaceutically acceptable salt thereof and water.The spironolactone can have a primary particle size distributioncharacterized by a D90 value of less than about 6 μm, less than about 1μm, or more preferably less than about 0.25 μm. The spironolactone canfurther have a primary particle size distribution characterized by a D50value of less than about 2.7 μm, less than about 0.75 μm, or morepreferably less than about 0.15 μm. The spironolactone can further havea primary particle size distribution characterized by a D10 value ofless than about 1.2 μm, less than about 0.50 μm, or more preferably lessthan about 0.10 μm.

In certain embodiments, the pharmaceutical composition comprises about0.10% w/w to about 7.5% w/w of spironolactone or a salt thereof. Incertain embodiments, the pharmaceutical composition can comprise fromabout 0.5% w/w to about 5% w/w of spironolactone or a salt thereof. Incertain embodiments, the pharmaceutical composition is an aqueoussuspension. In certain embodiments, the pharmaceutical compositionfurther comprises dioctyl sodium sulfosuccinate and/or hydroxyl propylcellulose. In certain embodiments, the pharmaceutical compositioncomprises about 0.01% w/w to about 1% w/w of dioctyl sodiumsulfosuccinate and/or about 0.01% to about 1.5% w/w of hydroxyl propylcellulose.

In certain embodiments of the present invention, a pharmaceuticalcomposition is provided consisting essentially of or consisting of atherapeutically effective amount of spironolactone or a pharmaceuticallyacceptable salt thereof, dioctyl sodium sulfosuccinate, hydroxyl propylcellulose, and water. The spironolactone can have a primary particlesize distribution characterized by a D90 value of less than about 6 μm,less than about 1 μm, or more preferably less than about 0.25 μm. Thespironolactone can further have a primary particle size distributioncharacterized by a D50 value of less than about 2.7 μm, less than about0.75 μm, or more preferably less than about 0.15 μm. The spironolactonecan further have a primary particle size distribution characterized by aD10 value of less than about 1.2 μm, less than about 0.50 μm, or morepreferably less than about 0.10 μm.

In certain embodiments, the pharmaceutical composition comprises about0.10% w/w to about 7.5% w/w of spironolactone or a salt thereof. Incertain embodiments, the pharmaceutical composition can comprise fromabout 0.5% w/w to about 5% w/w of spironolactone or a salt thereof. Incertain embodiments, the pharmaceutical composition is an aqueoussuspension. In certain embodiments, the pharmaceutical compositioncomprises about 0.01% w/w to about 1% w/w of dioctyl sodiumsulfosuccinate and about 0.01% to about 1.5% w/w of hydroxyl propylcellulose.

In certain embodiments, the pharmaceutical composition is capable ofdelivering spironolactone to the pilosebaceous unit of a patient. Incertain embodiments, the pharmaceutical composition is capable ofachieving dermal penetration of at least 1 mm in the patient, andpreferably about 2 or 3 mm in the subject.

In certain embodiments of the present invention, a method of treatingacne in a subject in need thereof is provided. In certain embodiments,the subject is a human male or female. In preferred embodiments, thesubject is a female human. The method comprises topically administeringto the subject the pharmaceutical compositions of spironolactonedescribed herein. In the methods of the present invention, thespironolactone or salt thereof can be delivered to the pilosebaceousunit. In the methods disclosed herein, administration of thepharmaceutical composition can result in dermal penetration ofspironolactone of at least 1 mm in the subject, and preferably about 2or 3 mm in the subject.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings, which are incorporated herein and form partof the disclosure, help illustrate various embodiments of the presentinvention and, together with the description, further serve to describethe invention to enable a person skilled in the pertinent art to makeand use the embodiments disclosed herein.

FIG. 1 shows a particle size distribution plot of SHR0302 in anexemplary pharmaceutical composition.

FIG. 2 shows a Fourier Transform Ion Cyclotron Resonance—HighResolution—Matrix Assisted Laser Desorption/Ionization Mass Spectrometry(FTICR-HR-MALDI) depth profile of an 0.3% SHR0302 topical 30% DMSO creamfor a first donor (Donor A).

FIG. 3 shows a FTICR-HR-MALDI depth profile of a 3% SHR0302 topicalsuspension in dimethicone for a first donor (Donor A).

FIG. 4 shows a FTICR-HR-MALDI depth profile of an 0.3% SHR0302 topical30% DMSO cream for a second donor (Donor B).

FIG. 5 shows a FTICR-HR-MALDI depth profile of a 3% SHR0302 topicalsuspension in dimethicone for a second donor (Donor B).

FIG. 6 shows a particle size distribution of spironolactone in asuspension of 5.0% spironolactone nano-milled in water with 0.05%dioctyl sulfosuccinate (DOSS) and 1.0% hydroxyl propyl cellulose.

FIG. 7 is a micrograph taken two weeks after storage past completion ofmilling for a 5% spironolactone suspension in cyclomethicone that wasroller milled to form a suspension having a D90 of less than about 5 μm.

FIG. 8 shows the cumulative amount of spironolactone appearing in thereceptor solution over 24 hours after a single 5.0 μl per cell (10 mgper cm² of skin tissue) for two exemplary formulations (Formulation 1and Formulation 2, described in Example 4) and a Comparative Gelformulation (also described in Example 4).

FIG. 9 shows the amount of spironolactone (ng) in the epidermis anddermis after 24 hours for two exemplary formulations (Formulation 1 andFormulation 2, described in Example 4) and a Comparative Gel formulation(also described in Example 4).

FIG. 10 is a micrograph taken two weeks after storage past completion ofmilling for a 5% spironolactone suspension in a 94.5:0.5 wt:wt blend ofcyclomethicone and dioctyl sodium sulfosuccinate that was roller milledto form a suspension having a D90 of less than about 5 μm.

FIG. 11 shows the cumulative amount of spironolactone appearing in thereceptor solution over 24 hours after a single 5.0 μl per cell (10 mgper cm² of skin tissue) for an exemplary aqueous suspension formulation(Formulation 1), Emulsion 1, Emulsion 2, and a Comparative Gelformulation (as described in further detail in Example 5).

FIG. 12 shows the amount of spironolactone (ng) in the epidermis anddermis after 24 hours for the exemplary aqueous suspension formulation(Formulation 1), Emulsion 1, Emulsion 2, and a Comparative Gelformulation (as described in Example 5).

FIG. 13 shows the average amount of spironolactone appearing in thereceptor solution over 24 hours after a single 5.0 μl per cell (10 mgper cm² of skin tissue) for eight exemplary formulations (S1-S7,described in Example 6, and Formulation 1, described in Example 4).

FIG. 14 shows the amount of spironolactone (ng) in the epidermis anddermis after 24 hours for eight exemplary formulations (S1-S7, describedin Example 6, and Formulation 1, described in Example 4).

DETAILED DESCRIPTION OF THE INVENTION

Before the present invention is described in detail below, it is to beunderstood that this invention is not limited to the particularmethodology, protocols, and reagents described herein as these may vary.It is also to be understood that the terminology used herein is for thepurpose of describing particular embodiments only, and is not intendedto limit the scope of the present invention which will be limited onlyby the appended claims. Unless defined otherwise, all technical andscientific terms used herein have the same meanings as commonlyunderstood by one of ordinary skill in the art to which this inventionbelongs.

All publications, patents and patent applications cited herein arehereby incorporated by reference in their entirety unless otherwisestated. Where the same term is defined in a publication, patent, orpatent application and the present disclosure incorporated herein byreference, the definition in the present disclosure represents acontrolling definition. For publications, patents and patentapplications referenced to describe a particular type of compound,chemistry, etc., the portion relating to such compounds, chemistry, etc.is the portion of the literature incorporated herein by reference.

Note that as used herein, the singular forms “a,” “an,” and “the”include plural referents unless the context clearly dictates otherwise.Thus, for example, “active ingredient” includes a single ingredient andtwo or more different ingredients and “sulfate salt” includes a singlesulfate salt as well as two or more different sulfate salts.

The term “about” when used in connection with a numerical value is meantto encompass numerical values within a range having a lower limit thatis 5% smaller than the indicated numerical value and having an upperlimit that is 5% larger than the indicated numerical value.

The term “effective” refers to an amount of a compound, agent,substance, formulation or composition that is of sufficient quantity toresult in a decrease in severity of disease symptoms, an increase infrequency and duration of disease symptom-free periods, or a preventionof impairment or disability due to the disease affliction. The amountmay be as a single dose or according to a multiple dose regimen, aloneor in combination with other compounds, agents or substances. One ofordinary skill in the art would be able to determine such amounts basedon such factors as a subject's size, the severity of a subject'ssymptoms, and the particular composition or route of administrationselected.

“Pharmaceutically acceptable” means generally safe for administration tohumans or animals. Preferably, a pharmaceutically acceptable componentis one that has been approved by a regulatory agency of the Federal or astate government or listed in the U.S. Pharmacopeia, published by theUnited States Pharmacopeial Convention, Inc., Rockville Md., or othergenerally recognized pharmacopeia for use in animals, and moreparticularly in humans.

A “pharmaceutical composition” according to the invention may be presentin the form of a composition, wherein the different active ingredientsand diluents and/or carriers are admixed with each other, or may takethe form of a combined preparation, where the active ingredients arepresent in partially or totally distinct form. An example for such acombination or combined preparation is a kit-of-parts.

As used herein, the terms “subject” “or patient” most preferably refersto a human being. The terms “subject” or “patient” may include anymammal that may benefit from the compounds described herein.

A “therapeutic amount” or “therapeutically effective amount” is anamount of a therapeutic agent sufficient to achieve the intendedpurpose. The effective amount of a given therapeutic agent will varywith factors such as the nature of the agent, the route ofadministration, the size of the subject to receive the therapeuticagent, and the purpose of the administration. The effective amount ineach individual case may be determined empirically by a skilled artisanaccording to established methods in the art.

The term “topical” with respect to administration of a drug orcomposition refers to the application of such drug or composition to theepithelial surface outside the body, including the skin or cornea. Forthis application, application to the inside of a body opening in whichthe mucosal surface does not contain pilosebaceous units, such as themouth, vagina or rectum is not considered a topical application.

As used herein, “treat,” “treating,” or “treatment” of a disease ordisorder means accomplishing one or more of the following: (a) reducingthe severity and/or duration of the disorder; (b) limiting or preventingdevelopment of symptoms characteristic of the disorder(s) being treated;(c) inhibiting worsening of symptoms characteristic of the disorder(s)being treated; (d) limiting or preventing recurrence of the disorder(s)in patients that have previously had the disorder(s); and (e) limitingor preventing recurrence of symptoms in patients that were previouslysymptomatic for the disorder(s).

The abbreviation “w/w” represents the relative concentration of thecomponents in the composition as “weight to weight” (i.e., percentagerefers to percentage of total weight), rather than based on volume orother quantities.

The present invention relates to pharmaceutical compositions for thetopical administration of a drug to the pilosebaceous unit and methodsfor administering the same. The inventors of the present invention havemade the surprising discovery that pharmaceutical compositionscomprising small particles of an active pharmaceutical ingredient can bedelivered to the pilosebaceous unit resulting in deeper penetration intothe dermis and improved efficacy. In preferred embodiments, thepharmaceutical composition comprises SHR0302 or spironolactone as anactive pharmaceutical ingredient.

In certain embodiments of the present invention, a pharmaceuticalcomposition is provided comprising a therapeutically effective amount ofan active pharmaceutical ingredient and a silicone selected from thegroup consisting of dimethicone, cyclomethicone, and combinationsthereof. In certain embodiments, the amount of active pharmaceuticalingredient can range from about 0.01% w/w to about 10% w/w, or fromabout 0.01% w/w to about 7.5% w/w, or from about 0.01% w/w to about 5%w/w, or from about 0.1% w/w to about 3% w/w. Exemplary ranges are fromabout 0.1% w/w to about 10% w/w, or from about 0.1% w/w to about 7.5%w/w, or from about 0.1% w/w to about 5% w/w, or from about 0.1% w/w toabout 5% w/w, or from about 1.0% w/w to about 5% w/w, or from about 0.3%w/w to about 5.0% w/w. For example, the pharmaceutical compositioncomprises any of the following w/w percents of active pharmaceuticalingredient: 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%,1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 7%, 1.8%, 1.9%, 1.0%, 2.1%, 2.2%,2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%,3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%,4.7%, 4.8%, 4.9%, 5.0% etc.

In the pharmaceutical compositions of the present invention, the activeingredient is present as small particles. Particle size of the drug canbe assessed using laser diffraction methods. Laser diffraction isrecognized by standards and guidance agencies including ISO and ASTM andis widely used to determine particle size distributions. In conductingthe assessment, the sample is passed through a laser beam, which resultsin laser light scattered at a range of angles. Detectors placed at fixedangles measure the intensity of light scattered at that position. Amathematical model is then applied to generate a particle sizedistribution. The particle size values reported herein are determined byusing a liquid or wet dispersion method.

In particle size determinations, the median value is defined as thevalue where half of the population resides above this point, and halfresides below this point. For particle size distributions the median iscalled the D50. The D50 is the size that splits the distribution withhalf above and half below this diameter. The distribution width may alsobe characterized by citing one, two or three values on the x-axis,typically some combination of the D10, D50, and D90. The D50, themedian, has been defined above as the diameter where half of thepopulation lies below this value. Similarly, 90 percent of thedistribution lies below the D90, and 10 percent of the population liesbelow the D10.

In certain embodiments, the active pharmaceutical ingredient has aprimary particle size distribution that is characterized by a D90 valueof less than about 20 μm, less than about 15 μm, less than about 10 μm,or more preferably less than about 5 μm. In certain embodiments, theactive pharmaceutical ingredient has a primary particle sizecharacterized by a D90 value of between about 0.001 μm, 0.01 μm, or 0.1μm and about 5 μm, 10 μm, 15 μm, and 20. The active pharmaceuticalingredient can further have a primary particle size distributioncharacterized by a D50 value of less than about 5 μm, less than about 2μm, less than about 1 μm, less than about 0.8 μm, or more preferablyless than about 0.7 μm. In certain embodiments, the activepharmaceutical ingredient has a primary particle size characterized by aD50 value of between about 0.001 μm, 0.01 μm, or 0.1 μm and about 0.7μm, 0.80 μm, 1.0 μm, 2.0 μm, or 5.0 μm. The active pharmaceuticalingredient can further have a primary particle size distributioncharacterized by a D10 value of less than about 1 μm, less than about0.5 μm, less than about 0.4 μm, or more preferably less than about 0.25μm. In certain embodiments, the active pharmaceutical ingredient has aprimary particle size characterized by a D10 value of between about0.0001 μm, 0.001 μm, or 0.01 μm and about 0.25 μm, 0.4 μm, 0.5 μm, or1.0 μm.

In certain embodiments, the pharmaceutical compositions of the presentinvention are capable of delivering the active pharmaceutical ingredientto the pilosebaceous unit. In certain embodiments, active pharmaceuticalingredient is capable of achieving dermal penetration of at least 1 mmin the subject.

In certain embodiments of the present invention, the pharmaceuticalcomposition comprises the JAK1 inhibitor,(3aR,5S,6aS)—N-(3-methoxyl-1,2,4-thiadiazol-5-yl)-5-(methyl(7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)hexahydrocyclopenta[c]pyrrole-2(1H)-carboxamide,which is also known as SHR0302 or ARQ-250. The terms SHR0302 and ARQ-250are used interchangeably herein. The structure of SHR0302 is:

SHR0302 is a potent small molecule inhibitor of JAK1 that has been shownto have a high selectively for JAK1 over JAK2, and thus has thepotential to treat inflammatory diseases without causing thehematopoietic adverse effects, such as anemia, thrombocytopenia, andneutropenia, associated with JAK2 inhibition. SHR0302 is disclosed inU.S. Pat. No. 9,527,851, which is hereby incorporated by reference.

In certain embodiments of the present invention, the pharmaceuticalcomposition comprises the aldosternone agonist,17-hydroxy-7α-mercapto-3-oxo-17α-pregn-4-ene-21-carboxylic acidγ-lactone acetate, which is also known as spironolactone. The structureof spironolactone is:

Spironolactone is a drug that acts at the mineralocorticoid receptorlevel by competitively inhibiting aldosterone binding. This steroidalcompound has been used for blocking aldosterone-dependent sodiumtransport in the distal tubule of the kidney in order to reduce edemaand to treat essential hypertension and primary hyperaldosteronism.Orally administrated spironolactone is also efficacious in the treatmentof women with acne. E. M. Attwa et al., Efficacy and safety of topicalspironolactone 5% gel versus placebo in the treatment of acne vulgaris,J. Dermatol. Venerol. 39:89-94 (2019); J. W. Charny et al.,Spironolactone for the treatment of acne in women, a retrospective studyof 110 patients, Int. J. Womens Dermatol. 3(2): 111-115 (2017).Spironolactone is commercially available under the tradenames ALDACTONE®and CAROSPIR®. Spironolactone is disclosed in U.S. Pat. No. 3,013,012,which is hereby incorporated by reference.

In the present invention, the pharmaceutical composition is administeredtopically. The pharmaceutical composition can include SHR0302 orspironolactone as a free base or a pharmaceutically acceptable salt.Suitable pharmaceutically acceptable salts can be found in Remington'sPharmaceutical Sciences, 17^(th) ed., Mack Publishing Company (1985),which is incorporated by reference herein.

In certain embodiments, the pharmaceutical composition comprises SHR0302having a primary particle size distribution characterized by a D90 valueof less than about 20 μm, less than about 15 μm, less than about 10 μm,or more preferably less than about 5 μm. In certain embodiments, theSHR0302 has a primary particle size characterized by a D90 value ofbetween about 0.01 μm, 0.1 μm, or 1.0 μm and about 5.0 μm, 10.0 μm, 15.0μm, or 20.0 μm.

The SHR0302 can further have a primary particle size distributioncharacterized by a D50 value of less than about 5 μm, less than about 2μm, less than about 1 μm, less than about 0.8 less, or more preferablyless than about 0.7 μm. In certain embodiments, the SHR0302 has aprimary particle size characterized by a D50 value of between about0.001 μm, 0.01 μm, or 0.1 μm and about 0.7 μm, 0.8 μm, 1.0 μm, 2.0 μm,or 5.0 μm.

The SHR0302 can further have a primary particle size distributioncharacterized by a D10 value of less than about 1 μm, less than about0.5 μm, less than about 0.4 μm, or more preferably less than about 0.25μm. In certain embodiments, the SHR0302 has a primary particle sizecharacterized by a D10 value of between about 0.0001 μm, 0.001 μm, or0.01 μm and about 0.25 μm, 0.4 μm, 0.5 μm, or 1.0 μm.

In certain embodiments, the pharmaceutical composition comprisesspironolactone having a primary particle size distribution characterizedby a D90 value of less than about 6 μm, less than about 5 μm, less thanabout 2 μm, less than about 1 μm, less than about 0.5 μm, less thanabout 0.25 μm, or more preferably less than about 0.2 μm. In certainembodiments, the spironolactone has a primary particle sizecharacterized by a D90 value of between about 0.001 μm, 0.01 μm, or 0.1μm and about 0.2 μm, 0.25 μm, 0.5 μm, 1 μm, 2 μm, 5 μm, or 6 μm.

The spironolactone can further have a primary particle size distributioncharacterized by a D50 value of less than about 2.7 μm, less than about2.0 μm, less than about 1.0 μm, less than about 0.75 μm, 0.5 μm, lessthan about 0.25 μm, less than about 0.2 μm, or more preferably less thanabout 0.15 μm. In certain embodiments, the spironolactone has a primaryparticle size characterized by a D50 value of between about 0.001 μm,0.01 μm, or 0.1 μm and about 0.15 μm, 0.2 μm, 0.25 μm, 0.5 μm, 0.75 μm,1.0 μm, 2.0 μm, or 2.7 μm.

The spironolactone can further have a primary particle size distributioncharacterized by a D10 value of less than about 1.2 μm, less than about1.0 μm, less than about 0.5 μm, less than about 0.25 μm, less than about0.15 μm, less than about 0.10 μm, or more preferably less than about0.08 μm. In certain embodiments, the spironolactone has a primaryparticle size characterized by a D10 value of between about 0.0001 μm,0.001 μm, or 0.01 μm and about 0.10 μm, 0.15 μm, 0.25 μm, 0.5 μm, 1.0μm, or 1.2 μm.

In certain embodiments, the pharmaceutical compositions further comprisea silicone selected from the group consisting of dimethicone,cyclomethicone, or combinations thereof. Dimethicone, also known aspolydimethylsiloxane (PDMS), is a polymeric organosilicon compound.Cyclomethicones are a group of methyl siloxanes, which unlikedimethicone, are cyclic organosilicon compounds. In certain embodiments,the pharmaceutical composition can comprise a combination of silicones,including dimethicone and cyclomethicone. For example, thepharmaceutical composition can comprisedimethicone-cyclomethicone-dimethicone/vinyl dimethicone. Additionalmethyl siloxane compatible excipients such as cyclopentasiloxane,dimethiconol and phenyl trimethicone may be added to dimethicone and/orcyclomethicone, to adjust aesthetics or viscosity. The silicone, such asdimethicone or cyclomethicone has a polarity similar to sebum, allowingfor the pharmaceutical compositions to target follicular delivery.

In certain embodiments, the pharmaceutical composition is a suspension,wherein the active ingredient (e.g., SHR0302 or spironolactone) issuspended in the silicone (e.g., dimethicone or cyclomethicone). Incertain embodiments, the pharmaceutical compositions can be formulatedas an emulsion. For example, the pharmaceutical composition can beformulated as one of the following forms:

An oil-in-water emulsion: The product may be an emulsion comprising adiscrete phase of a hydrophobic component and a continuous aqueous phasethat includes water and optionally one or more polar hydrophilicexcipients as well as solvents, co-solvents, salts, surfactants,emulsifiers, and other components. These emulsions may includewater-soluble or water-swellable polymers that help to stabilize theemulsion.

A water-in-oil emulsion: The compositions may be an emulsion thatincludes a continuous phase of a hydrophobic component and an aqueousphase that includes water and optionally one or more polar hydrophiliccarrier(s) as well as salts or other components. These emulsions mayinclude oil-soluble or oil-swellable polymers as well as one or moreemulsifier(s) to help to stabilize the emulsion.

A microemulsion: These are clear, thermodynamically stable isotropicliquid systems that contain oil, water and surfactants, frequently incombination with a cosurfactant. Microemulsions may be water continuous,oil continuous or bicontinuous mixtures. The formulations may optionallyalso contain water up to 60% by weight. Higher levels may be suitable insome compositions.

A nanoemulsion: These are isotropic dispersed systems that containwater, oil, and an emulsifier. The system may be an oily systemdispersed in an aqueous system, or an aqueous system dispersed in anoily system forming droplets or oily phases of nanometric sizes.Nanoemulsions often have higher loading capacity for lipophilic activeingredients than microemulsions. Hydrophobic and hydrophilic activeingredients can also be formulated in nanoemulsion. Nanoemulsions may beformed by any suitable method known in the art, including high-pressurehomogenization, microfluidization, and phase-inversion temperature.

In certain embodiments, the pharmaceutical composition consistsessentially of the active ingredient and a silicone selected from thegroup consisting of dimethicone, cyclomethicone, or combinationsthereof. Alternatively, the pharmaceutical compositions may beformulated with additional components, including conventionally found incosmetic and pharmaceutical topical products.

In certain embodiments, the pharmaceutical composition is an suspension,wherein the active ingredient (e.g., spironolactone) is suspended inwater. In certain embodiments, the pharmaceutical composition comprisesbetween about 90% and 99% w/w water. In certain embodiments, thepharmaceutical composition consists essentially of or consists ofspironolactone, dioctyl sodium sulfosuccinate, hydroxyl propylcellulose, and water. In certain embodiments, the pharmaceuticalcomposition does not comprise an additional thickening agent or apreservative. The inclusion of additional pharmaceutical excipients,such as an additional thickening agent or a preservative, can inactivatethe ability of the suspended spironolactone to target the pilosebaceousunit.

In certain embodiments, the pharmaceutical composition may be formulatedwith additional components, including conventionally found in cosmeticand pharmaceutical topical products. In certain embodiments, theadditional components comprise no more than 3%, 2%, 1%, or 0.5% w/w ofthe pharmaceutical composition. In certain embodiments, thepharmaceutical composition further comprises dioctyl sodiumsulfosuccinate. In certain embodiments, the pharmaceutical compositionfurther comprises hydroxyl propyl cellulose. In certain embodiments, thepharmaceutical composition comprises about 0.01% w/w to about 1% w/w ofdioctyl sodium sulfosuccinate and/or about 0.01% to about 1.5% w/w ofhydroxyl propyl cellulose.

In certain embodiments, the pharmaceutical composition further comprisesone or more of hydroxypropylmethyl cellulose (HPMC),Polyvinylpyrrolidone (PVP K30), poloxamers such as Poloxamer 407, orpolysorbates such as polysorbate 80. In certain embodiments, thepharmaceutical composition comprises about 0.1% to about 1.5% HPMC. Incertain embodiments, the pharmaceutical composition comprises about 0.1%to about 1.5% PVP K30. In certain embodiments, the pharmaceuticalcomposition comprises about 0.1% to about 1.5% Poloxamer 407. In certainembodiments, the pharmaceutical composition comprises about 0.1% toabout 1.5% Polysorbate 80. In certain embodiments, the pharmaceuticalcomposition comprises a pharmaceutically acceptable cellulose polymersuch as methyl cellulose (MC), ethyl cellulose (EC), hydroxyethylcellulose (HEC), carboxymethyl cellulose (CMC), sodium carboxymethylcellulose (NaCMC), or microcrystalline cellulose. In certainembodiments, the pharmaceutical composition comprises about 0.1% toabout 1.5% of a cellulose polymer.

Surfactants

In certain embodiments, the pharmaceutical composition may include oneor more surfactants or co-surfactants. Surfactants include, but are notlimited to short-chain alcohols, alkane diols and triols, alkylphosphate esters, polyethylene glycols and glycol ethers, polyethylenestearyl ethers, including those sold under the tradenames Brij S2, BrijS20, Brij 721, Brij 38, Brij 52, Brij 56, and Brij W1, pyrrolidinederivatives, bile salts, sorbitan fatty acid esters and polyoxyethylenesorbitan fatty acid esters. In preferred embodiments, the surfactant isdioctyl sodium sulfosuccinate.

Polymers and Thickeners

In certain embodiments, the pharmaceutical composition may includeinsoluble organic polymeric thickeners such as natural and syntheticpolymers or inorganic thickeners such as acrylates copolymer, carbomer1382, carbomer copolymer type B, carbomer homopolymer type A, carbomerhomopolymer type B, carbomer homopolymer type C, acrylamide/sodiumacryloyldimethyl taurate copolymer, carboxy vinyl copolymer,carboxymethylcellulose, carboxypolymethylene, carrageenan, guar gum,xanthan gum, hydroxyethyl cellulose, hydroxypropyl cellulose,microcrystalline wax, and methylcellulose.

Additional Components

In certain embodiments, the pharmaceutical composition may includeadditional components such as carriers and excipients conventionallyfound in cosmetic and pharmaceutical topical products. Additionalcomponents including but not limited to antioxidants (e.g., BHT, BHA,ascorbic acid, tocopherol, citric acid, propyl gallate, sodiummetabisulfite), sequestering agents, stabilizers, buffers, pH adjustingagents (preferably agents which result in an acidic pH, including butnot limited to gluconolatone, citric acid, lactic acid, and alphahydroxyacids), skin penetration enhancers, skin protectants (includingbut not limited to petrolatum, paraffin wax, dimethicone, glycerylmonoisostearate, isopropyl isostearate, isostearyl isostearate, cetylalcohol, potassium cetyl phosphate, cetyl behenate and behenic acid),chelating agents, suspending agents (e.g., xanthan gum), dyes, pigments,diluents, fragrances, and other excipients to improve the stability oraesthetics, may be added to the composition.

Administration and Dosage

The present invention includes methods of treating hair loss conditions,such as alopecia, androgenic hair loss, hypothrichosis, and telogeneffluvium. The methods can include treating a hair loss condition in apatient in need thereof by administering to the patient the compositionsof SHR0302 or sprinolactone described herein.

In preferred embodiments, the present invention includes methods oftreating alopecia areata (AA). AA is one of the most highly prevalentautoimmune diseases, leading to hair loss due to the collapse of immuneprivilege of the hair follicle and subsequent autoimmune destruction. AAis a skin disease that leads to hair loss on the scalp and elsewhere.Prior to the present invention, topical administration of JAK inhibitorshave not shown reproducible clinical efficacy. Without being bound bytheory, the inability of JAK inhibitors to treat AA prior to the claimedinvention is believed to be due to insufficient drug delivery to thepilosebaceous unit, and more specifically, the hair bulb. The inventorsof the present invention have made the surprising discovery that thepharmaceutical compositions disclosed herein are capable of penetratingthe at least about 1 mm, at least about 2 mm, and at least about 3 mminto the hair follicle of an AA patient.

In certain embodiments, the present invention provides methods fortreating AA in a patient in need thereof, comprising topically applyinga therapeutically effective amount of the SHR0302 pharmaceuticalcompositions described herein to the patient. In certain embodiments,the active ingredient, SHR0302, can be administered in a therapeuticallyeffective amount. In certain embodiments, the amount of SHR0302 canrange from about 0.01% w/w to about 7.5% w/w, or from about 0.01% w/w toabout 5% w/w, or from about 0.1% w/w to about 3% w/w. Exemplary rangesare from about 0.01% w/w to about 5% w/w, or from about 0.01% w/w toabout 3% w/w, or from about 0.1% w/w to about 3% w/w, or from about 0.3%w/w to about 3.0% w/w. For example, the topical formulation comprisesany of the following w/w percents of SHR0302: 0.1%, 0.2%, 0.3%, 0.4%,0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%,7%, 1.8%, 1.9%, 1.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%,2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%,4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, etc.

The present invention further provides methods of treating acne in apatient in need thereof. Acne is a disorder of the pilosebaceous unitslocated on the face, chest and back. The acne can be one selected fromthe group consisting of acne, acne vulgaris, inflammatory acne,non-inflammatory acne, acne fulminans, nodular papulopustular acne, acneconglobata, acne rosacea, rosacea, acne excoriee, adult-onset acne,persistent-recurrent acne past teenaged years, and acne associated withother disorders. In certain embodiments, the patient is a human male orfemale patient. In preferred embodiments, the patient is a human female.Further, the patient can be: (a) experiencing acne flares that cyclewith menstruation; (b) inflicted with adult onset acne orpersistent-recurrent acne past teenage years, even in the absence ofclinical or laboratory signs of hyperandrogenism, (c) on oralcontraceptives and exhibiting moderate to serve acne, especially with ahormonal pattern clinically; or (d) not responding to conventionaltherapy and who are not candidates for oral isotretinoin therapy.

In certain embodiments, the present invention provides methods fortreating acne in a patient in need thereof, comprising topicallyapplying a therapeutically effective amount of the spironolactonepharmaceutical compositions described herein to the patient. In certainembodiments, the active ingredient, spironolactone, can be administeredin a therapeutically effective amount. In certain embodiments, theamount of spironolactone can range from about 0.01% w/w to about 10%w/w, or from about 0.01% w/w to about 7.5% w/w, or from about 0.01% w/wto about 5% w/w, or from about 0.1% w/w to about 3% w/w. Exemplaryranges are from about 0.1% w/w to about 10% w/w, or from about 0.1% w/wto about 7.5% w/w, or from about 0.1% w/w to about 5% w/w, or from about0.1% w/w to about 3% w/w, or from about 1.0% w/w to about 5% w/w, orfrom about 0.3% w/w to about 5.0% w/w. For example, the topicalformulation comprises any of the following w/w percents ofspironolactone: 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%,1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 7%, 1.8%, 1.9%, 1.0%, 2.1%,2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%,3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%,4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%,5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%,7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, etc.

In certain embodiments, the pharmaceutical composition is administeredtopically as a regimen, such as at regular intervals. For example, atopical pharmaceutical composition can be administered once daily, twicedaily, thrice daily, once per week, twice per week, three times perweek, or four times per week. The pharmaceutical compositions can beadministered for a prescribed period of time. For example, a topicalpharmaceutical composition can be administered for a period of about twoweeks to at least about six months, or until an improvement in skincondition or disease is visually observed. Exemplary periods of time forthe treatment regimen include two weeks, one month, six weeks, twomonths, three months, four months, five months, six months, sevenmonths, eight months, nine months, or one year. In preferredembodiments, the topical pharmaceutical composition is administeredtwice or thrice daily for a period of at least 3 months, 4 months, 5months, or 6 months.

The following examples illustrate certain embodiments of the inventionwithout limitation.

EXAMPLES

While various embodiments have been described above, it should beunderstood that they have been presented by way of example only, and notlimitation. Thus, the breadth and scope of the present disclosure shouldnot be limited by any of the above-described exemplary embodiments.Moreover, any combination of the above-described elements in allpossible variations thereof is encompassed by the disclosure unlessotherwise indicated herein or otherwise clearly contradicted by context.

Comparative Example 1

Comparative Example 1 was prepared as a 0.3% SHR0302 topical creamhaving the composition set forth in Table 1.

TABLE 1 Composition of SHR0302 Cream SHR0302 0.3% Ingredient CreamSHR0302 0.30% w/w Dimethyl Sulfoxide (DMSO) 30.0% w/w Propylene Glycol15% w/w Polyethylene Glycol 200 15% w/w Cyclomethicone 7.0% w/wST-Elastomer 10 2.0% w/w Dimethicone 1.0% w/w Pemulen TR 1 0.8% w/wCarbopol 974P 1.5% w/w EDTA 0.05% w/w BHT 0.05% w/w Benzyl Alcohol 2.0%w/w D-Limonene 0.1% w/w Trolamine (25% solution to q.s. ad pH 5.5 adjustpH) Purified Water q.s. ad 100% Total 100%

Example 1

A pharmaceutical composition comprising 3% SHR0302 suspended indimethicone was prepared. The SHR0302 used in the composition had aparticle size distribution as set forth in FIG. 1 . The primary particlesize distribution of the SHR0302 is characterized by a D10 value of lessthan about 0.25 μm; a D50 value of less than about 0.7 μm; and a D90value of less than about 5 μm. The pharmaceutical composition was atransparent solution due to the particles being too small to scattervisible light. A Malvern Metasizer Model 3000 using the Hydro MVdispersion unit was used to determine the particle size distributionprofile of SHR0302. The sample preparation procedure performed was asfollows: weigh 10-20 mg of SCP processed ARQ-250 into a 30-mL vial, add20 mL of ethyl acetate, sonicate the suspension with an ultrasonic probeat 40% power for 2 minutes in a 5° C. water bath, and transfer thesample suspension to the Malvern Hydro MV dispersion unit to obtainobscuration between 5 and 15%. The instrument parameters were: (i)Refractive Index of Particles: 1.5; (ii) Refractive Index of Dispersant1.395; (iii) Absorption Index: 0.01; (iv) Measurement Duration: 10seconds; (v) Number of Measurements: 3; (vi) Stir Speed: 3500 rpm; (vii)Ultrasonics: Off.

Example 2

The ability of Comparative Example 1 and Example 1 to penetrate intohuman cadaver scalp skin was assessed. Two different human cadaver scalpskins (Donor A and B) were mounted on special tension cell. A dose (7.5μL) of either Comparative Example 1 or Example 1 was applied to thescalp skin specimen for 6 hours. All formulation residue was washed fromthe skin. An 8 mm punch biopsy was taken from the dermis side of theskin and flash frozen in liquid nitrogen. Serial 10 μm cryosections weretaken, wherein an hematoxylin and eosin (H&E) stain was prepared forevery other section, and adjacent sections were retained for analysisusing Fourier Transform Ion Cyclotron Resonance—High Resolution13 MatrixAssisted Laser Desorption/Ionization Mass Spectrometry (FTICR-HR-MALDI).The FTICR-HR-MALDI analysis was performed using a Bruker 7TFTICR-HR-MALDI MS system.

FIGS. 2 and 4 show a FTICR-HR-MALDI depth profile of an 0.3% SHR0302topical 30% DMSO cream for Donor A and Donor B, respectively. As shownin FIG. 2 , Comparative Example 1 achieved less than 160 μm of maximumdermal penetration in Donor A. As shown in FIG. 4 , Comparative Example1 achieved less than 500 μm of maximum dermal penetration in Donor B.The results are consistent with a pharmaceutical composition capable ofdelivering drug across the stratum corneum and indicate that drug is notpenetrating below the upper capillary plexus.

FIGS. 3 and 5 show a FTICR-HR-MALDI depth profile of a 3% SHR0302topical suspension in dimethicone for Donor A for Donor A and Donor B,respectively. As shown in FIGS. 3 and 5 , Example 1 achieved greaterthan 1 mm of maximum dermal penetration. The results surprisingly showthat the pharmaceutical composition is capable of delivering drug to thehair bulb.

Example 3

A 50 mg/mL aqueous suspension of 5% spironolactone containing 0.5%dioctyl sodium sulfosuccinate and 1% hydroxyl propyl cellulose wassuccessfully nano-milled to provide stable submicron particles ofsuspension of drug particles after storage for two weeks at 5° C. andambient light. The spironolactone had a particle size distribution asset forth in FIG. 6 . A Horiba Laser Scattering Particle SizeDistribution Analyzer Model LA-950 was used to determine the volumebased distribution profile of spironolactone. Circulation, agitation,and ultrasound were all turned off and the instrument was set to manualiteration mode.

A stable 0.3% spironolactone oil-in-water emulsion was prepared havingthe composition set forth in Table 2:

TABLE 2 Composition of Spironolactone Oil-in-Water EmulsionSpironolactone Oil-in-Water Ingredient Emulsion Spironolactone 0.30% w/wCyclomethicone 10.0% w/w Methylparaben 0.10% w/w Propylbaraben 0.01% w/wSepineo P600 4.0% w/w Purified Water q.s. ad 100% Total 100%

Example 4

A 5.0%-5.5% spironolactone suspension in water containing 0.05%-0.055%dioctyl sodium sulfosuccinate and 1.0-1.1% hydroxyl propyl cellulose wasnano-milled to achieve the particle size distribution shown in FIG. 6 .The composition of the finished product suspension is listed in Table 3as Formulation 1. A 5% spironolactone suspension in cyclomethicone wasroller milled to form a suspension having a D90 of less than about 5 μmas shown in FIG. 7 (microphotograph taken after two weeks of storagepast completion of milling). The composition of this finished productsuspension is listed in Table 3 as Formulation 2. A comparative gelformulation described in the literature was prepared and is listed inTable 3 as Comparative Gel. (Attwa E M, Ibrahim A M, Abd El-Halim M F,Mahmoud H M, Efficacy and safety of topical spironolactone 5% gel versusplacebo in the treatment of acne vulgaris, Egypt J Dermatol Venerol(2019); 39:89-94.)

TABLE 3 Composition of Two Deep Dermal Drug Delivery Formulations and aComparative Gel from the Literature. Formulation Formulation Comparative1 2 Gel Ingredient (% w/w) (% w/w) (% w/w) Spironolactone 5.0-5.5 5.05.0 Ethanol — — 20.0 Glycerin — — 10.0 Propylene glycol — — 10.0 Lacticacid — — 5.0 Methyl cellulose — — 3.0 Sodium benzoate — — 0.03Cylclomethicone — 95.0 — Dioctyl sodium sulfosuccinate  0.05-0.055 — —Hydroxyl propyl 1.0-1.1 — — cellulose Water q.s. ad 100% — 46.97

In vitro skin penetration testing (IVPT) was used to determine howrapidly the different formulations crossed excised human skin. Humancadaver skin was procured from two donors (Caucasian female age=48abdomen skin dermatomed to an average thickness of 580 μm and Hispanicmale age=50 abdomen skin dermatomed to an average thickness of 910 μm).Dermatomed skin was received frozen from a US tissue bank and stored at−20° C. until use. Skin was loaded onto vertical Franz cells having a0.503 cm² (8 mm in diameter) diffusion area and a receptor chamberfilled with 3.0 ml of 4% BSA in water containing 0.01% gentamicinsulfate thermostated at 32° C. Using a positive displacement pipette, 5microliters of each formulation was dosed on each Franz Cell (10 mg persquare centimeter of skin). Receptor solutions were analyzed using avalidated LC-MS/MS (Kinetex C18, 5 μm, 2.1×50 mm column, ShimadzuLC20ADXR pumps and AB Sciex API 4000 Turbo Spray detector). Thecumulative amount of spironolactone assayed in the receptor solution isthe average of four replicate IVPT measurements.

To determine the levels of spironolactone retained in the epidermis anddermis 24-hours after dosing the skin, the skin surface was cleaned ofany unabsorbed and unpenetrated spironolactone. This was accomplished bywiping the tissue surface with a Q-tip wetted with 1×PBS three timesfollowed by two tape strippings. The epidermis (including the stratumcorneum) was removed from the dermis and soaked in 4.0 ml of aDMSO/Acetonitrile (ACN) (50/50 v/v) mixture for overnight at roomtemperature using an orbit shaker. The remaining dermis layer was cutinto small pieces and extracted with 4.0 ml of the DMSO/ACN mixture forovernight at room temperature using an orbit shaker. Extracts of thedermis and epidermis were analyzed using a validated LC-MS/MS (KinetexC18, 5 μm, 2.1×50 mm column, Shimadzu LC20ADXR pumps and AB Sciex API4000 Turbo Spray detector).

FIG. 8 illustrates the cumulative amount of spironolactone appearing inthe receptor solution over 24 hours after a single 5.0 μl per cell (10mg per cm² of skin tissue) for Formulation 1, Formulation 2, and theComparative Gel. In FIG. 8 , each plotted value is the average of fourseparate pieces of excised human skin. FIG. 9 illustrates the amount ofspironolactone (ng) in the epidermis and dermis after 24 hours forFormulation 1, Formulation 2, and the Comparative Gel. As seen in FIG. 8the comparative gel with 5% dissolved spironolactone delivered morespironolactone across excised human skin than either the 5%spironolactone water suspension or the 5% spironolactone cyclomethiconesuspension. However, significantly greater deposition of spironolactoneinto the epidermis (location of the infundibulum of the pilosebaceousunit) for both suspensions is seen in FIG. 9 . High levels of API in theepidermis and dermis indicates that spironolactone targets thepilosebaceous unit and has significantly greater follicular depositionfrom the aqueous suspension (D90<0.5 μm which is Formulation 1) andcyclomethicone suspension (D90<5.0 μm which is Formulation 2).

Example 5

A 5.0%-5.5% spironolactone suspension in water containing 0.05%-0.055%dioctyl sodium sulfosuccinate and 1.0-1.1% hydroxyl propyl cellulose wasnano-milled to achieve the particle size distribution shown in FIG. 6 .The composition of the finished product suspension is listed in Table 1as Formulation 1. An emulsion was formed by mixing the excipientsdisclosed in Table 1 as Emulsion 1. SEPINEO™ P 600 is a thickening,emulsifying and stabilizing polymer Acrylamide/Sodium AcryloyldimethylTaurate Copolymer/Isohexadecane & Polysorbate 80 distributed by SeppicInc., Fairfield N.J. Table 4 lists the final composition of Emulsion 1in which the cyclomethicone oil phase was a 5% spironolactone suspensionin cyclomethicone was roller milled to form a suspension having a D90 ofless than about 5 μm as shown in FIG. 7 (microphotograph taken after twoweeks of storage past completion of milling). Table 4 also lists thefinal composition of Emulsion 2 in which the cyclomethicone oil phasewas 5% spironolactone suspension in a 94.5:0.5 wt:wt blend ofcyclomethicone and dioctyl sodium sulfosuccinate that was roller milledto form a suspension having a D90 of less than about 5 μm as shown inFIG. 10 (microphotograph taken after two weeks of storage pastcompletion of milling). A comparative gel formulation described in theliterature was prepared and is listed in Table 4 as Comparative Gel.(Attwa EM, Ibrahim AM, Abd El-Halim MF, Mahmoud HM, Efficacy and safetyof topical spironolactone 5% gel versus placebo in the treatment of acnevulgaris, Egypt J Dermatol Venerol (2019); 39:89-94.)

TABLE 4 Composition of Deep Dermal Drug Delivery Formulations and aComparative Gel from the Literature. Formulation Emulsion EmulsionCompar- 1 1 2 ative Gel Ingredient (% w/w) (% w/w) (% w/w) (% w/w)Spironolactone 5.0-5.5 4.8 4.8 5.0 Ethanol — — — 20.0 Glycerin — — —10.0 Propylene glycol — — — 10.0 Lactic acid — — — 5.0 Methyl cellulose— — — 3.0 Sodium benzoate — — — 0.03 Cylclomethicone — 9.5 9.45 —Dioctyl sodium  0.05-0.055 0.043 0.048 — sulfosuccinate Hydroxyl propyl1.0-1.1 0.86 0.86 — cellulose Sepineo P600 — 4.0 4.0 — Water q.s. adq.s. ad q.s. ad 46.97 100% 100% 100%

In vitro skin penetration testing (IVPT) was used to determine howrapidly the different formulations crossed excised human skin. Humancadaver skin was procured from two donors (Caucasian female age=48abdomen skin dermatomed to an average thickness of 580 μm and Hispanicmale age=50 abdomen skin dermatomed to an average thickness of 910 μm).Dermatomed skin was received frozen from a US tissue bank and stored at−20° C. until use. Skin was loaded onto vertical Franz cells having a0.503 cm² (8 mm in diameter) diffusion area and a receptor chamberfilled with 3.0 ml of 4% BSA in water containing 0.01% gentamicinsulfate thermostated at 32° C. Using a positive displacement pipette, 5microliters of each formulation was dosed on each Franz Cell (10 mg persquare centimeter of skin). Receptor solutions were analyzed using avalidated LC-MS/MS (Kinetex C18, 5 μm, 2.1×50 mm column, ShimadzuLC20ADXR pumps and AB Sciex API 4000 Turbo Spray detector). Thecumulative amount of spironolactone assayed in the receptor solution isthe average of four replicate IVPT measurements.

To determine the levels of spironolactone retained in the epidermis anddermis 24-hours after dosing the skin, the skin surface was cleaned ofany unabsorbed and unpenetrated spironolactone. This was accomplished bywiping the tissue surface with a Q-tip wetted with 1× PBS three timesfollowed by two tape strippings. The epidermis (including the stratumcorneum) was removed from the dermis and soaked in 4.0 ml of aDMSO/Acetonitrile (ACN) (50/50 v/v) mixture for overnight at roomtemperature using an orbit shaker. The remaining dermis layer was cutinto small pieces and extracted with 4.0 ml of the DMSO/ACN mixture forovernight at room temperature using an orbit shaker. Extracts of thedermis and epidermis were analyzed using a validated LC-MS/MS (KinetexC18, 5 μm, 2.1×50 mm column, Shimadzu LC20ADXR pumps and AB Sciex API4000 Turbo Spray detector).

FIG. 11 illustrates the cumulative amount of spironolactone appearing inthe receptor solution over 24 hours after a single 5.0 μl per cell (10mg per cm² of skin tissue) for Formulation 1, Emulsion 1, Emulsion 2,and the Comparative Gel. In FIG. 11 , each plotted value is the averageof four separate pieces of excised human skin. FIG. 12 illustrates theamount of spironolactone (ng) in the epidermis and dermis after 24 hoursfor Formulation 1, Emulsion 1, Emulsion 2, and the Comparative Gel. Asseen in FIG. 12 the comparative gel with 5% dissolved spironolactonedelivered more spironolactone across excised human skin than the 5%spironolactone water suspension or either of the two emulsionformulations. However, significantly greater deposition ofspironolactone into the epidermis (location of the infundibulum of thepilosebaceous unit) for the aqueous suspension is seen in FIG. 12compared to the comparative gel having dissolved spironolactone or thesilicone oil emulsions having submicron particles of suspendedspironolactone. High levels of API in the epidermis and dermis indicatesthat spironolactone targets the pilosebaceous unit and has significantlygreater follicular deposition from the aqueous suspension (D90<0.5 μmwhich is Formulation 1).

Example 6

A 5.0%-5.5% spironolactone suspension in water containing 0.05%-0.055%dioctyl sodium sulfosuccinate and 1.0-1.1% hydroxyl propyl cellulose wasnano-milled to achieve the particle size distribution shown in FIG. 6 .To this aqueous suspension different preservatives, thickening agentsand ethanol were added to this aqueous suspension as shown in Table 5.Seven different formulations (S1-S7) were prepared as set forth in Table5.

TABLE 5 Composition of the Comparative Spironolactone (S) AqueousSuspension Formulations. Formulation S-1 S-2 S-3 S-4 S-5 S-6 S-7Components (% w/w) (% w/w) (% w/w) (% w/w) (% w/w) (% w/w) (% w/w)Spironolactone 4.98 4.94 4.94 4.90 4.89 5.00 4.93 Methylparaben 0.100.10 0.10 — 0.10 — 0.10 Propylparaben 0.01 0.01 0.01 — 0.01 — 0.01Phenoxyethanol — — — 1.00 — — — Benzyl Alcohol — — — — — 1.00 — Ethanol— — — — — 10.00  — Polysorbate 80 — — — — 1.00 — — Poloxamer 407 — — — —— — 0.20 Carbopol 974 0.30 — — — — — — Hydroxypropyl — 1.00 — 1.00 1.001.00 1.00 cellulose Hyaluronic — — 1.00 — — — — Acid Water q.s. ad q.s.ad q.s. ad q.s. ad q.s. ad q.s. ad q.s. ad containing 100% 100% 100%100% 100% 100% 100% 0.05-0.055% dioctyl sodium sulfosuccinate and1.0-1.1% hydroxyl propyl cellulose

In vitro skin penetration testing (IVPT) was used to determine howrapidly the different formulations crossed excised human skin. Humancadaver skin was procured from two donors (Caucasian female age=44abdomen skin dermatomed to an average thickness of 710 μm and Caucasianfemale age=48 abdomen skin dermatomed to an average thickness of 578μm). Dermatomed skin was received frozen from a US tissue bank andstored at −20° C. until use. Skin was loaded onto vertical Franz cellshaving a 0.503 cm² (8 mm in diameter) diffusion area and a receptorchamber filled with 3.0 ml of 4% BSA in water containing 0.01%gentamicin sulfate thermostated at 32° C. Using a positive displacementpipette, 5 microliters of each formulation was dosed on each Franz Cell(10 mg per square centimeter of skin). Receptor solutions were analyzedusing a validated LC-MS/MS (Kinetex C18, 5 μm, 2.1×50 mm column,Shimadzu LC20ADXR pumps and AB Sciex API 4000 Turbo Spray detector). Thecumulative amount of spironolactone assayed in the receptor solution isthe average of four replicate IVPT measurements.

To determine the levels of spironolactone retained in the epidermis anddermis 24-hours after dosing the skin, the skin surface was cleaned ofany unabsorbed and unpenetrated spironolactone. This was accomplished bywiping the tissue surface with a Q-tip wetted with 1× PBS three timesfollowed by two tape strippings. The epidermis (including the stratumcorneum) was removed from the dermis and soaked in 4.0 ml of aDMSO/Acetonitrile (ACN) (50/50 v/v) mixture for overnight at roomtemperature using an orbit shaker. The remaining dermis layer was cutinto small pieces and extracted with 4.0 ml of the DMSO/ACN mixture forovernight at room temperature using an orbit shaker. Extracts of thedermis and epidermis were analyzed using a validated LC-MS/MS (KinetexC18, 5 μm, 2.1×50 mm column, Shimadzu LC20ADXR pumps and AB Sciex API4000 Turbo Spray detector).

FIG. 13 illustrates the average amount of spironolactone appearing inthe receptor solution over 24 hours after a single 5.0 μl per cell (10mg per cm² of skin tissue) for S1-S7 and Formulation 1 (as described inExample 4). In FIG. 13 , each plotted value is the average of fourseparate pieces of excised human skin. FIG. 14 illustrates the amount ofspironolactone (ng) in the epidermis and dermis after 24 hours for S1-S7and Formulation 1 (as described in Example 4). FIG. 14 shows that therewas significantly greater deposition of spironolactone in the epidermisand dermis after topical dosing with an aqueous suspension(Formulation 1) compared with the seven other formulations (S1-S7)having the same particle size distribution set forth in FIG. 6 . Thedata indicates that even minor modifications of adding a preservative orgelling agent inactivates the ability of suspended spironolactone totarget the pilosebaceous unit.

The foregoing description has been presented for purposes ofillustration and description. This description is not intended to limitthe invention to the precise form disclosed. Persons of ordinary skillin the art will appreciate that modifications and substitutions of thebasic inventive description may be made.

What is claimed is:
 1. A pharmaceutical composition comprising: atherapeutically effective amount of spironolactone or a pharmaceuticallyacceptable salt thereof, wherein a primary particle size distribution ofthe spironolactone is characterized by a D90 value of less than about 6μm; and a silicone selected from the group consisting of dimethicone andcyclomethicone.
 2. The pharmaceutical composition of claim 1, whereinthe composition is an oil-in-water emulsion.
 3. The pharmaceuticalcomposition of claim 1, wherein the composition comprises about 0.10%w/w to about 7.5% w/w of spironolactone or a salt thereof.
 4. Thepharmaceutical composition of claim 1, wherein a primary particle sizedistribution of the spironolactone is characterized by a D90 value ofless than about 1 μm.
 5. The pharmaceutical composition of claim 1,wherein a primary particle size distribution of the spironolactone ischaracterized by a D90 value of less than about 0.25 μm.
 6. Thepharmaceutical composition of claim 1, wherein a primary particle sizedistribution of the spironolactone is characterized by a D50 value ofless than about 2.7 μm.
 7. The pharmaceutical composition of claim 1,wherein a primary particle size distribution of the spironolactone ischaracterized by a D10 value of less than about 1.2 μm.
 8. A method oftreating acne in a subject in need thereof comprising: topicallyadministering to the subject a pharmaceutical composition comprising:(a) a therapeutically effective amount of spironolactone or apharmaceutically acceptable salt thereof, wherein a primary particlesize distribution of the spironolactone is characterized by a D90 valueof less than about 6 μm; and (b) a silicone selected from the groupconsisting of dimethicone and cyclomethicone.
 9. The method of claim 8,wherein the spironolactone is delivered to the pilosebaceous unit. 10.The method of claim 8, wherein the spironolactone achieves dermalpenetration of at least 1 mm in the subject.
 11. The method of claim 8,wherein the pharmaceutical composition is an oil-in-water emulsion. 12.The method of claim 8, wherein the composition comprises about 0.10% w/wto about 7.5% w/w of spironolactone or a salt thereof.
 13. The method ofclaim 8, wherein a primary particle size distribution of thespironolactone is characterized by a D90 value of less than about 1 μm.14. The method of claim 8, wherein a primary particle size distributionof the spironolactone is characterized by a D90 value of less than about0.25 μm.
 15. The method of claim 8, wherein a primary particle sizedistribution of the spironolactone is characterized by a D50 value ofless than about 2.7 μm.
 16. The method of claim 8, wherein a primaryparticle size distribution of the spironolactone is characterized by aD10 value of less than about 1.2 μm.